Abstract
AbstractThe advancement of fluorescence microscopy techniques has opened up new opportunities for visualizing proteins and unraveling their functions in living biological systems. Small‐molecule organic dyes, which possess exceptional photophysical properties, small size, and high photostability, serve as powerful fluorescent reporters in protein imaging. However, achieving high‐contrast live‐cell labeling of target proteins with conventional organic dyes remains a considerable challenge in bioimaging and biosensing due to their inadequate cell permeability and high background signal. Over the past decade, a novel generation of fluorogenic and cell‐permeable dyes has been developed, which have substantially improved live‐cell protein labeling by fine‐tuning the reversible equilibrium between a cell‐permeable, nonfluorescent spirocyclic state (unbound) and a fluorescent zwitterion (protein‐bound) of rhodamines. In this review, we present the mechanism and design strategies of these fluorogenic and cell‐permeable rhodamines, as well as their applications in bioimaging and biosensing.
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